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The two cysteines may be separated from each other by many amino acids in the primary sequence of a polypeptide, or may even be located on two different polypeptide chains; the folding of the polypeptide chain s brings the cysteine residues into proximity, and permits covalent bonding of their side chains. In the absence of an enzyme, only a small proportion of a population of molecules may possess enough energy to achieve the transition state between reactant and product. Prolyl residue H HN C. The rate of an enzyme-catalyzed reaction increases with substrate concentration until a maximal velocity Vmax is reached Figure 5. However, as seen in the RBC, the presence of 2,3-BPG significantly reduces the affinity of hemoglobin for oxygen, shifting the oxygen dissociation curve to the right Figure 3.